Broad cross-reactive epitopes of the H5N1 influenza virus identified by murine antibodies against the A/Vietnam/1194/2004 hemagglutinin

Mie Kobayashi-Ishihara, Hitoshi Takahashi, Kazuo Ohnishi, Kengo Nishimura, Kazutaka Terahara, Manabu Ato, Shigeyuki Itamura, Tsutomu Kageyama, Yasuko Tsunetsugu-Yokota

Research output: Contribution to journalArticle

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Abstract

There is an urgent need for a rapid diagnostic system to detect the H5 subtype of the influenza A virus. We previously developed monoclonal antibodies (mAbs) against the H5 hemagglutinin (HA) for use in a rapid diagnostic kit. In this study, we determined the epitopes of the anti-H5 HA murine mAbs OM-b, AY-2C2, and YH-1A1. Binding assays of the mAbs to different strains of H5 HAs indicated that OM-b and AY-2C2 cross-reacted with HAs from clades 1, 2.1.3.2, 2.2, and 2.3.4, whereas YH-1A1 failed to bind to those of clades 2.1.3.2 and 2.3.4. HA chimeras revealed that the epitopes for each of the mAbs were in the HA1 region. Analysis of escape mutants revealed that OM-b and AY-2C2 mAbs interacted mainly with amino acid residues D43 and G46, and the YH-1A1 mAb interacted with G139 and K or R140 of H5 HA. Multiple alignments of H5 HA protein sequences showed that D43 and G46 were very conserved among H5N1 HAs, except those in clade 2.2.1 and clade 7 (88.7%). The epitope for YH-1A1 mAb was highly variable in the HAs of H5N1, although it was well conserved in those of H5N2-N9. The OM-b and AY-2C2 mAbs could bind to the HAs of clades 1.1 and 2.3.2.1 that are currently epidemic in Asia, and we conclude that these would be effective for the detection of H5N1 infections in this region.

Original languageEnglish
Article numbere99201
JournalPloS one
Volume9
Issue number6
DOIs
Publication statusPublished - 2014 Jun 19
Externally publishedYes

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H5N1 Subtype Influenza A Virus
Vietnam
Hemagglutinins
hemagglutinins
Orthomyxoviridae
Viruses
epitopes
Epitopes
monoclonal antibodies
Monoclonal Antibodies
antibodies
Antibodies
mice
analytical kits
chimerism
Influenza A virus
Assays
amino acid sequences
Amino Acids
mutants

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Broad cross-reactive epitopes of the H5N1 influenza virus identified by murine antibodies against the A/Vietnam/1194/2004 hemagglutinin. / Kobayashi-Ishihara, Mie; Takahashi, Hitoshi; Ohnishi, Kazuo; Nishimura, Kengo; Terahara, Kazutaka; Ato, Manabu; Itamura, Shigeyuki; Kageyama, Tsutomu; Tsunetsugu-Yokota, Yasuko.

In: PloS one, Vol. 9, No. 6, e99201, 19.06.2014.

Research output: Contribution to journalArticle

Kobayashi-Ishihara, M, Takahashi, H, Ohnishi, K, Nishimura, K, Terahara, K, Ato, M, Itamura, S, Kageyama, T & Tsunetsugu-Yokota, Y 2014, 'Broad cross-reactive epitopes of the H5N1 influenza virus identified by murine antibodies against the A/Vietnam/1194/2004 hemagglutinin', PloS one, vol. 9, no. 6, e99201. https://doi.org/10.1371/journal.pone.0099201
Kobayashi-Ishihara, Mie ; Takahashi, Hitoshi ; Ohnishi, Kazuo ; Nishimura, Kengo ; Terahara, Kazutaka ; Ato, Manabu ; Itamura, Shigeyuki ; Kageyama, Tsutomu ; Tsunetsugu-Yokota, Yasuko. / Broad cross-reactive epitopes of the H5N1 influenza virus identified by murine antibodies against the A/Vietnam/1194/2004 hemagglutinin. In: PloS one. 2014 ; Vol. 9, No. 6.
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